ABSTRACTIn this study, novel nickel oxide (NiO) flowers like nanostructures were fabricated onto gold coated glass substrate by hydrothermal method using high alkaline pH medium. The structural study of nickel oxide nanostructures was performed by scanning electron microscopy (SEM) and X-ray differaction (XRD) techniques. Nickel oxide nanostructures are highly dense, uniform and possess good crystalline quality. The so prepared structures were investigated for their electrochemical properties by cyclic voltammetry and amperometric techniques. The nickel oxide flower like morphology has shown good electrochemical performances for the oxidation of glucose. The presented sensing material was able to detected glucose in a wide range of concentration of 0.001 mM to 8 mM with a high sensitivity (123μmA/mM) and regression coefficient of 0.99. Moreover, the NiO nanostructures based sensor is highly reproducible, stable, exhibiting a fast response time and selective in the response. All the obtained results indicate the potential use of this material in the development of enzyme free sensors for the detection of glucose.